Claudin 1 antibody

The resected terminal ileum tissues were mixed with the lysis buffer (1 % Triton X-100; 50 mmol/L Tris–HCl, pH 7.6; 150 mmol/L NaCl; and 1 % protease inhibitor) and centrifuged at 12,000 r/min for 5 min at 4 °C. The supernatant was separated and collected. Caco-2 cells were harvested, washed with the cold PBS (PH 7.4) and mixed with the lysis buffer. The total proteins of tissues and Caco-2 cells were exacted with the protein extraction kit (Nanjing KeyGEN Biotech Co. Ltd., Nanjing, China) following the manufacturer’s instructions. The protein concentration was determined with the bicinchoninic acid (BCA) protein assay. After separated with SDS-PAGE, the proteins were transferred to a PVDF membrane. The PVDF membrane was block with 5 % skim milk in Tris-buffered saline containing 0.05 % Tween-20 (TTBS), incubated with the primary antibodies (occludin antibody (Santa Cruz, CA, USA, 1:1000 dilution), claudin-1 antibody (Abcam, Cambridge, MA, USA, 1:1000 dilution) and beta-actin antibody (Zhongshan Goldenbridge Biological Technology, Beijing, China, 1:1000 dilution)) overnight at 4 °C and subsequently incubated with the secondary antibody for 2 h at 37 °C. The bands were photographed and quantified with the ChemiDoc XRS documentation system (Bio-Rad Laboratories). The intensity of the beta-actin band was as the internal reference.

DSS (AppliChem, Lot#：A3261‐0250 Germany); the six medicinal herbs of QBD raw powder, as shown in Table 1 (Pharmacy Department, The Second Hospital affiliated to Shanxi University of TCM, Xianyang, China) were decoting up 150 mL by traditional decocting method for enema； Mesalazine Enemas (V Vifor AG Zweigniederlassung Medichemie Ettingen， H20150127, Switzerland); faecal occult blood (FOB) (Baso Diagnostics Inc， Zhuhai， china); NF‐κB p65 antibody (Cell Signaling, #8242, Danvers, MA, USA); Phospho‐NF‐κB p65 antibody (Cell Signaling, #3039); p44/42 MAPK (ERK1/2) (antibody (Cell Signaling，#4695); Phospho‐p44/42 MAPK (ERK1/2) antibody (Cell Signaling, #4370); MMP‐9 antibody (Merck, AB19016, Burlington, MA, USA); Ki67 antibody (Merck, AB9260); Cleaved Caspase‐3 antibody (Cell Signaling, #9664); Caspase‐3 antibody (Cell Signaling, #9662); β‐Actin antibody (Cell Signaling,# 4970) Notch1 antibody (Cell Signaling, #3608s); ZO‐1 antibody (abcam, ab96587, Cambridge, MA, USA); Occludin antibody (abcam, ab 21632); claudin‐1 antibody (abcam, ab15098); primer synthesis (Takara Bio Inc, Dalian, China).